I'm working on camelina seed sterilization and have some doubts about the process. I followed a protocol used by my lab mate, but I'm unsure about a couple of things:
Here's what I did for camelina seed sterilization:
- I placed the seeds in a 5% bleach solution for about 10 minutes because I was slow in removing the bleach solution from the tubes using a pipette.
- Then, I added 90% ethanol for 30 seconds, but due to my slow work, the ethanol remained in the tube for more than a minute.
- I washed the seeds with 10 ml of sterile water only once and dried them on filter paper for 15 minutes.
- Finally, I transferred the seeds to already prepared MS plates supplemented with Kanamycin, which is my selectable marker.
Could you let me know if my approach is correct and if the concentrations and durations I used are safe for camelina seeds?
Will my camelina seeds grow?
Also i stored the remaining dried sterilized seeds in another tubes. How can i use it again ?
Try to germinate the seeds to see if it works. It can then be included in your results! :)
- Bleach Concentration: Consider using 5% bleach for 5-10 minutes if you observe any adverse effects with 10%.
- Ethanol Concentration: Switch to 70% ethanol for 30 seconds.
- Washing: Increase the number of sterile water washes to 2-3 times.
- Timeliness: Try to be more precise with the timing to avoid overexposure to sterilizing agents.
By making these adjustments, you can optimize the sterilization process for camelina seeds and improve the likelihood of successful germination and growth.
The most effective method to sterilize Camelina seeds is surface sterilization using a combination of chemical and physical treatments. A common protocol involves:
This process eliminates surface contaminants without damaging the seed's viability. The exact concentrations and durations may be adjusted based on seed batch and specific research requirements. It's crucial to perform this procedure under sterile conditions to prevent recontamination. For particularly sensitive applications, additional steps like hot water treatment or the use of hydrogen peroxide may be considered.